The genomic region encompassing the L protein gene and a small open reading frame (ORF 6) of Rice yellow stunt virus (RYSV) has been sequenced, thus completing the nucleotide sequence of the RYSV genome. The genome organization of RYSV is unique in the rhabdoviruses because it contains two additional genes when compared to the basic gene order of the family Rhabdoviridae. Phylogenetic analysis revealed that the amino acid sequence of the RYSV L protein is most closely related to that of the L protein of Sonchus yellows net virus, another nucleorhabdovirus. However, the RYSV L protein has a unique acidic N-terminal domain distinct from that of other rhabdoviruses. Moreover, the polypeptide encoded by the ORF 6 was detected by immunoblot analysis in purified RYSV virions. Thus RYSV provides the first example in the family Rhabdoviridae that a small ORF between the G and L genes encodes a virion protein.Rice yellow stunt virus (RYSV), synonymous with Rice transitory yellowing virus, is a species in the genus Nucleorhabdovirus of the Rhabdoviridae (Walker et al., 2000). RYSV has a non-segmented, negative-sense singlestranded RNA genome. We have previously sequenced the RYSV genomic RNA from cDNA clones representing the 39 leader (Wang et al., 1999), the nucleocapsid protein (N) (Fang et al., 1994), the phosphoprotein (P, formerly designated non-structural protein) (Zhu et al., 1997), a protein (P3) of unknown function , the matrix protein (M) and the glycoprotein (G) , as well as the 59 trailer region (Wang et al., 1999). In this paper, we report the sequence of the genomic region between the glycoprotein gene and the 59 trailer, thus completing the sequence of the entire RYSV genome (14 042 nucleotides). In this region, a large open reading frame (ORF) encoding the polymerase (L) and a small ORF (ORF 6) capable of encoding 93 amino acids (aa) were found on the viral complementary (vc) strand. Furthermore, we present evidence that the polypeptide encoded by ORF 6, named P6, is a virion-associated protein. Thus RYSV encodes seven genes and has, among the characterized rhabdoviruses, the unique genome organization 39 leader-N-P-3-M-G-6-L-59 trailer. In comparison to the basic gene order represented by the genome of Vesicular stomatitis virus (VSV), the RYSV genome has two additional genes: gene 3 and gene 6. Gene 3 is located between the P and M genes where additional ORFs were identified in the genomes of all plant rhabdoviruses examined so far [Sonchus yellows net virus (SYNV) (Scholthof et al., 1994), Lettuce necrotic yellows virus (LNYV) (Wetzel et al., 1994) and Northern cereal mosaic virus (NCMV) (Tanno et al., 2000)] and the insect rhabdovirus Sigma virus (SigmaV) (Teninges et al., 1993). However, in the position between the G and L genes where the RYSV gene 6 is interposed, the presence of the extra gene(s) has only been described for some animal rhabdoviruses [Adelaide River virus (ARV) , Bovine ephemeral fever virus (BEFV) (McWilliam et al., 1997), Rabies virus (RABV) (Tordo et al., 1986), Infectious haemat...